CAJ | 학술논문

目的研究乙型肝炎病毒C蛋白在NK细胞中的表达及其对NK细胞功能的影响.方法用脂质体转染法将HBV C基因真核表达载体pHBI-CMV-HBC转入NK-92细胞,通过WesternBlot检测C基因在NK-92细胞中表达,用ELISA法检测NK细胞分泌IFN-γ水平,MTT法检测NK细胞对HepG2细胞的细胞毒作用.结果 Western Blot证实转染有pHBI-CMV-HBC的NK-92细胞能表达HBV C蛋白.转染了重组真核表达质粒的NK-92细胞IFN-γ水平均高于空质粒对照组和空白对照组(P<0.01).与两对照组相比,重组真核表达质粒转染组NK-92细胞对于HepG2细胞的细胞毒活性明显提高,差异具有统计学意义(P<0.01).结论 HBc基因瞬时高表达可影响NK-92细胞分泌IFN-γ和细胞毒功能.
목적 연구을형간염병독C단백재NK세포중적표체급기대NK세포공능적영향.방법 용지질체전염법장HBV C기인진핵표체재체pHBI-CMV-HBC전입NK-92세포,통과WesternBlot검측C기인재NK-92세포중표체,용ELISA법검측NK세포분비IFN-γ수평,MTT법검측NK세포대HepG2세포적세포독작용.결과 Western Blot증실전염유pHBI-CMV-HBC적NK-92세포능표체HBV C단백.전염료중조진핵표체질립적NK-92세포IFN-γ수평균고우공질립대조조화공백대조조(P<0.01).여량대조조상비,중조진핵표체질립전염조NK-92세포대우HepG2세포적세포독활성명현제고,차이구유통계학의의(P<0.01).결론 HBc기인순시고표체가영향NK-92세포분비IFN-γ화세포독공능.
Objective To investigate the influence of hepatitis B virus C protein on the function of natural killer cell. Methods Recombinant eukaryotic expression plasmid pHB1-CMV-HBC was constructed and confirmed by double restrictive enzyme digestion and DNA sequencing analysis. Then the recombinant plasmid was transfected into NK-92 cells with lipofectamine encapsuled. The transfected NK-92 cells containing expressive HBV C protein was confirmed by Western Blot analysis. ELISA was employed to determine the IFN-γ level secreted by NK-92 cells. And finally the cytotoxicities of NK cells were analysed by MrIT colorimetry, with the hepatoblastoma cell line (HepG2) as target cell. Results Western blotting confirmed the expression of HBV C protein in the NK-92 cells transfected with pHBI-CMV-HBC. NK cytotoxicities and IFN-γ secretion level of NK-92 cells transfected with recombinant plasmid significantly increased compared to control NK-92 ceils transfected with blank plasmid (P< 0.01) and untransfected NK-92 cells(P<0.01). Conclussion Transient expression of HBC can increase IFN-γ secretion and cytotoxicities of NK-92 cells.